Matching class I and class II human leukocyte antigens (HLA) is a critical component of successful bone marrow transplantation. There is an urgent need for a quick, accurate, easy to use, high throughput method for molecular HLA typing, however current methods are limited in one or more of these area. To address this problem, Pel-Freez Clinical Systems has been investigating a novel technology called reference strand mediated heteroduplex analysis (RSCA). To adequately meet the market demand and realize the full potential of this technology, automation is required. The proposed automated RSCA HLA typing system would offer many advantages over existing methods, including higher resolution, higher throughput, lower cost per typing, and less human intervention. To establish feasibility for development of such a system. Phase I studies will focus on regent development for HLA-A and -B loci, construction of a prototype automated system utilizing the GeneSys GTI-9600 genetic analyzer, and validating the instrument. Phase II will focus on complete automation from blood sample to allele assignment and chemistry development for other HLA loci. In phase III, Pel-Freez Clinical Systems will commercialize the system and various reagents kits. PROPOSED COMMERCIAL APPLICATION: The resulting system will provide state-of-the-art high throughput instrumentation for HLA typing which can be applied to other molecular diagnostic applications such as identification of bacterial strains or HIV variants.